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Micro-satellite DNA (Deoxyribose Nucleic Acid) method for identifying reproductive mode of termite nest group

A technology of microsatellites and microsatellite sites, which is applied in the field of microsatellite DNA to identify the reproductive mode of termite nests, can solve the problems of not knowing the biological characteristics, and achieve the effect of improving pertinence

Inactive Publication Date: 2012-07-11
HUAZHONG AGRI UNIV
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AI Technical Summary

Problems solved by technology

However, due to the hidden nature of termite nesting and foraging, many important biological characteristics of termites in the natural environment are still not grasped: such as the type, number and relationship of reproductive ants in termite nests? What is the ratio of the nest colony controlled by the original king ant queen to the nest colony controlled by the supplementary reproductive ants in a certain area?

Method used

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Embodiment Construction

[0016] The invention consists of seven key steps including sample collection, genome DNA extraction, microsatellite site search, primer design and optimization, genotype determination, chi-square test and reproductive mode determination.

[0017] The present invention is described in detail below through specific examples.

[0018] Sample collection: collect termite samples from 10 nest colonies of S. blackbreast termites in Changsha, collect 25 worker ants for each nest colony, and the distance between two sampling points should be more than 20m. After sampling, put worker ants into sample tubes, soak them in 95% alcohol, label them, and store them in a -20°C refrigerator for later use. During the sampling process, the longitude, latitude and altitude of each sampling point were measured with a handheld GPS locator.

[0019] Genomic DNA extraction: extract the genomic DNA of all samples according to the method of the existing animal tissue genomic DNA extraction kit. Use ND...

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Abstract

The invention relates to a micro-satellite DNA (Deoxyribose Nucleic Acid) method for identifying a reproductive mode of a termite nest group, which comprises the following steps of: collecting above 20 termite samples from a same nest group in the open air, extracting a sample genome DNA and diluting; searching a micro-satellite site related to a termite reproductive mode on GenBank, designing a primer on the side wing of a micro-satellite repetitive sequence by utilizing software Primer Premier 5.0, carrying out electrophoresis on an automatic analyzer after the termite samples DNA are subjected to PCR (Polymerase Chain Reaction) amplification by selecting a micro-satellite primer and determining the genotype of each strip corresponding sample in a electrophoretic diagram by using SAGAGT software; and carrying out chi-square test on observed genotype frequencies and expected genotype frequencies of the termite samples based on a Mendel's law and judging the reproductive mode of the termite nest group. The micro-satellite DNA identification for reproductive modes of ten reticulitermes chinensis nest groups in Changsha by adopting the micro-satellite DNA method shows that the reticulitermes chinensis nest groups include seven simple nest groups, two amplified nest groups and one mixed nest group. The micro-satellite DNA method has the advantages that the operation is simple and convenient, the results are reliable, the reproductive modes of the termite group can be accurately judged, and the important reference value is provided for the formulation of a termite prevention strategy.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, in particular to a microsatellite DNA method for identifying the reproductive mode of termite nests. Background technique [0002] Microsatellite DNA, also known as simple sequence repeat (SSR), has the advantages of high level of heterozygosity, mutation rate, and parental inheritance mode, and has been widely used in gene mapping, germplasm resource identification, and genetic relationship analysis. , genetic map construction, species identification, and evolutionary analysis (see "Chromosoma", 2000, 109: 365-371). [0003] Termites are the oldest social insects, which seriously damage houses, reservoirs and dams, and agricultural and forestry plants. Due to the hidden life of termites, complex nest structure and diverse reproductive modes, it is difficult for people to directly observe the habits of termites, so termite control work is very blind. In the 1950s, Chinese scholars beg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 黄求应李刚华雷朝亮
Owner HUAZHONG AGRI UNIV
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